CAJ | 학술논문

根据实验室高通量测序所得核转录因子κB（ NF-κB） NF-κB1、 NF-κB2和I-κBα部分序列，应用荧光定量PCR技术，检测虹彩病毒（ Rana grylio virus，RGV）胁迫下NF-κB1、NF-κB2以及I-κBαmRNA在东北林蛙（ Rana dybowskii）肝脏、脾脏、皮肤中的差异表达，并对表达差异进行统计学分析。结果显示：在东北林蛙不同组织中，NF-κB1和NF-κB2的表达水平有差异，NF-κB1表达量约是NF-κB2的2倍，且皮肤和脾脏中的表达量远高于肝脏。在虹彩病毒刺激初，2个基因在3个组织中表达量立即下调，受刺激0．5 h后2个亚基在3个组织中都有上调趋势；肝脏和皮肤中，在刺激后2 h（在肝脏中）和4 h（在皮肤中）快速上调至峰值；在脾脏中，2亚基上调趋势缓慢，分别在16 h（NF-κB2）和24 h（NF-κB1）达到峰值；2亚基表达至峰值后开始下调，并在8 h（在肝脏中）和12 h（在皮肤中）下调至初始水平，在脾脏中2亚基72 h以后才达到初始水平。 I-κBα在受刺激初期也立即下调而后上调，上调至峰值的时间点与NF-κB基本相同。统计分析结果表明：各实验组基因表达量与对照组存在统计学差异（P＜0．05）。
근거실험실고통량측서소득핵전록인자κB（ NF-κB） NF-κB1、 NF-κB2화I-κBα부분서렬，응용형광정량PCR기술，검측홍채병독（ Rana grylio virus，RGV）협박하NF-κB1、NF-κB2이급I-κBαmRNA재동북림와（ Rana dybowskii）간장、비장、피부중적차이표체，병대표체차이진행통계학분석。결과현시：재동북림와불동조직중，NF-κB1화NF-κB2적표체수평유차이，NF-κB1표체량약시NF-κB2적2배，차피부화비장중적표체량원고우간장。재홍채병독자격초，2개기인재3개조직중표체량립즉하조，수자격0．5 h후2개아기재3개조직중도유상조추세；간장화피부중，재자격후2 h（재간장중）화4 h（재피부중）쾌속상조지봉치；재비장중，2아기상조추세완만，분별재16 h（NF-κB2）화24 h（NF-κB1）체도봉치；2아기표체지봉치후개시하조，병재8 h（재간장중）화12 h（재피부중）하조지초시수평，재비장중2아기72 h이후재체도초시수평。 I-κBα재수자격초기야립즉하조이후상조，상조지봉치적시간점여NF-κB기본상동。통계분석결과표명：각실험조기인표체량여대조조존재통계학차이（P＜0．05）。
By high-throughput sequencing , the partial sequence of NF-κB1、NF-κB2 are two subunits of nuclear factorκB ( NF-κB) and I-κBα.Under Rana grylio virus stress, we used real-time PCR to quantitatively detect NF-κB1, NF-κB2 and I-κBαmRNA in skin, liver and spleen of Rana dybowskii and statistically analyzed.the expression difference .The expression levels of the NF-κB1 and NF-κB2 have differences in different organizations , and NF-κB1 expression level is two times as high as NF-κB2.The expression level in skin and spleen is much higher than that in the liver.Inthe early stimulating stage, the expression of NF-κB1 and NF-κB2 is down-regulated immediately in three organizations.After 0.5 hour, two subunit expressions begin to increase.Expression rises quickly to the peak in the liver and skin-coerced for2 hours (in the liver) or 4 hours ( in the skin) later.In the spleen, upward trend turns slow, and the maximum is at 16 hours ( NF-κB2) or 24 hours (N F-κB1).After reaching itspeak , the expressions of two subunits declineagain and they drop to the initial level at 8 hours (in the liver) or 12 hours (in the skin), respectively.In the spleen, the initial level occurs after 72 hours.Ex-pression of I-κBαis down-regulated immediately in the early stimulating.The raised time is substantially identical with that of NF-κB.By statistical analysis, there is statistically significant difference in the expression levels between experimental group and control group (P <0.05).